Compositions and Methods for Treating Multiple Sclerosis and Related Disorders

ABSTRACT

The present disclosure is in the field of pharmaceutical compositions suitable for the treatment of diseases in mammals. The disclosure provides novel compositions comprising non-pathogenic fecal microbes for treating multiple sclerosis and related diseases. The disclosure also provides methods for treating a subject with the compositions disclosed herein.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/406,749 filed Oct. 11, 2016, the contents of which are hereinincorporated by reference in their entirety.

FIELD

The present disclosure relates to pharmaceutical compositions andmethods suitable for treating multiple sclerosis.

BACKGROUND

Mammals harbor diverse microbial species in their gastrointestinal (GI)tracts. Interactions between these microbes and between microbes and thehost, e.g. the host immune system, shape a microbiota. A healthymicrobiota provides the host with multiple benefits, includingcolonization resistance to a broad spectrum of pathogens, essentialnutrient biosynthesis and absorption, and immune stimulation thatmaintains a healthy gut epithelium and an appropriately controlledsystemic immunity. An unbalanced microbiota (also called ‘dysbiosis’ ordisrupted symbiosis) may lose its function and results in increasedsusceptibility to pathogens, altered metabolic profiles, or induction ofproinflammatory signals that can lead to local or systemic inflammationor autoimmunity. Additionally, such a disrupted microbiota may beinfected by incoming pathogen or pathogens, which can cause pain,diarrhea, gas, and constipation among other symptoms. Hence, theintestinal microbiota plays a significant role in the pathogenesis ofmany disorders such as pathogenic infections of the gut.

Implantation or administration of human colonic microbiota into thebowel of a sick patient is called Fecal Microbiota Transplantation(FMT), also commonly known as fecal bacteriotherapy. FMT is believed torepopulate the gut with a diverse array of microbes that control keypathogens by creating an ecological environment inimical to theirproliferation and survival. It represents a therapeutic protocol thatallows a fast reconstitution of a normal compositional and functionalgut microbial community.

FMT has been used to treat Clostridium difficile infection (CDI). FMThas also been suggested in treating other gut infective agents such asE. coli and Vancomycin resistant Enterococci (VRE). It entails infusionsthrough a colonoscope, an enema or via a nasojejunal tube of humanmicrobiota either in the form of homogenised stool, or cultured stoolcomponents such as Clostridia, to implant in the colon and therebydisplace or eradicate pathogenic bacteria, e.g., C. difficile.

Without being bound to any theory, multiple sclerosis (MS) is consideredby some as a disabling autoimmune disease of the central nervous system.In multiple sclerosis the immune system attacks the protective myelinsheath of nerve fibers. Damage to the myelin sheath slows down or stopsnerve signaling and causes miscommunication between the brain and spinalcord and other parts of the body. The disease can progress to permanentdamage of the nerves. Multiple sclerosis consists of four disease types,including clinically isolated syndrome (CIS), relapsing-remittingmultiple sclerosis (RRMS), primary progressive multiple sclerosis(PPMS), and secondary progressive multiple sclerosis (SPMS). Symptoms ofmultiple sclerosis vary depending on the amount of nerve damage,location of damage and severity of the disease. Attacks can last fordays, weeks, or months, followed by periods of remission. People whosuffer from severe multiple sclerosis may lose the ability to walk.Other symptoms can include loss of balance, muscle spasms, numbness,motor difficulties, tremors and limb weakness. Additional symptoms canaffect the bowel or bladder. These include constipation and stoolleakage, difficulty urinating, frequent urination, and urine leakage. Asubject with multiple sclerosis can also suffer from double vision, eyediscomfort, rapid eye movements and vision loss. Multiple sclerosis mayfurther cause memory loss, difficulty solving problems, depression,hearing loss, slurred speech and difficulty with chewing and swallowing.

Existing treatments for multiple sclerosis involve medication that candecrease or diminish symptoms and slow down the disease. Multiplesclerosis affects women more than men and is most commonly diagnosedbetween ages 20 to 40. The hospitalization rates of multiple sclerosispatients decreased by 75% in 2011, in comparison to 1984, however,hospitalization rates of multiple sclerosis patients remain higher thanhospitalization rates of the general population. See, Marrie et al.,Neurology 2014; 83: 929-937. Thus, there is a need for more effectivetreatments for multiple sclerosis that are easier to administer.

SUMMARY

The present disclosure provides compositions, methods, and dosingregimens for treating multiple sclerosis or preventing multiplesclerosis relapse.

In one aspect, the present disclosure provides a method for treatingmultiple sclerosis in a subject in need thereof, where the methodcomprises administering to the subject a pharmaceutically active dose ofa therapeutic composition comprising or derived frpm live non-pathogenicfecal bacteria or a sterile fecal filtrate. In one aspect, a sterilefecal filtrate originates from a donor stool. In another aspect, asterile fecal filtrate originates from cultured microorganisms.

In another aspect, this disclosure provides use of a compositioncomprising live non-pathogenic fecal bacteria in the manufacture of amedication for the treatment of multiple sclerosis.

In one aspect, a method provided here is for treating a form of multiplesclerosis selected from the group consisting of clinically isolatedsyndrome (CIS), relapsing-remitting multiple sclerosis (RRMS), primaryprogressive multiple sclerosis (PPMS), and secondary progressivemultiple sclerosis (SPMS).

In one aspect, the present disclosure provides a method for treatingmultiple sclerosis in a subject in need thereof, where the methodcomprises administering orally to the subject a pharmaceutically activedose of a therapeutic composition comprising or derived from live,non-pathogenic, synthetic bacterial mixture or live, non-pathogenic,purified or extracted, fecal microbiota, where the dose is administeredat a dosing schedule of at least once or twice daily or at least once ortwice weekly for at least three, eight, ten, or twenty consecutiveweeks. In a further aspect, the dose is administered at a dosingschedule of at least once or twice daily or at least once or twiceweekly for at least four, five, six, seven, eleven, twelve, thirteen,fourteen, fifteen, sixteen, seventeen, eighteen, or nineteen consecutiveweeks.

In one aspect, the present disclosure provides a method for treatingmultiple sclerosis in a subject in need thereof, where the methodcomprises administering orally to the subject a pharmaceutically activedose of a therapeutic composition comprising a liquid, frozen,lyophilized, or encapsulated sterile fecal filtrate, where the dose isadministered at a dosing schedule of at least once or twice daily or atleast once or twice weekly for at least three, eight, ten, or twentyconsecutive weeks.

In one aspect, a method achieves a remission, cure, response, orresolution rate of multiple sclerosis of at least about 80%.

In an aspect, a fecal microbiota in a therapeutic composition comprisesa donor's substantially entire and non-selected fecal microbiota,reconstituted fecal material, or synthetic fecal material.

DETAILED DESCRIPTION

Unless defined otherwise herein, terms are to be understood according toconventional usage by those of ordinary skill in the relevant art.

As used herein, the term “treating” refers to (i) completely orpartially inhibiting a disease, disorder or condition, for example,arresting its development; (ii) completely or partially relieving adisease, disorder or condition, for example, causing regression of thedisease, disorder and/or condition; or (iii) completely or partiallypreventing a disease, disorder or condition from occurring in a patientthat may be predisposed to the disease, disorder and/or condition, buthas not yet been diagnosed as having it. Similarly, “treatment” refersto both therapeutic treatment and prophylactic or preventative measures.

As used herein, “therapeutically effective amount” or “pharmaceuticallyactive dose” refers to an amount of a composition which is effective intreating the named disease, disorder or condition.

As used herein, “microbiota,” and “flora” refer to a community ofmicrobes that live in or on a subject's body, both sustainably andtransiently, including eukaryotes, archaea, bacteria, and viruses(including bacterial viruses (i.e., phage)). A non-selected fecalmicrobiota refers to a community or mixture of fecal microbes derivedfrom a donor's fecal sample without selection and substantiallyresembling microbial constituents and population structure found in suchfecal sample.

As used herein, a “sterile fecal filtrate” or a “non-cellular fecalfiltrate” refers to a liquid component of a fecal material, where theliquid component is free or substantially free of cell-based livingorganisms (e.g., bacteria, fungi, or their spores), but retainsbacteriophages and non-cellular biological materials. Preferably, anon-cellular or sterile fecal filtrate is also free of viruses foreukaryotic host cells.

As used herein, “remission, cure, or resolution rate” refers to thepercentage of patients that are cured or obtain remission or completeresolution of a condition in response to a given treatment.Quantitatively, a patient responds to a treatment positively when thepatient's MSIS-29 score decreases by at least 2 from baseline to week 8.Remission, cure, or resolution of multiple sclerosis refers to reducedor no sign of symptoms selected from the group consisting ofconstipation and stool leakage, difficulty urinating, frequenturination, urine leakage, bladder infection, double vision, eyediscomfort, rapid eye movements, vision loss, memory loss, difficultysolving problems, depression, hearing loss, slurred speech, difficultywith chewing or swallowing, dizziness, vertigo, fatigue, itching, pain,sexual discomfort, tremors, walking difficulties, or muscle spasms.

As used herein, “response rate” refers to the percentage of patientsthat respond positively (e.g., reduced severity or frequency of one ormore symptoms) to a given treatment. A multiple sclerosis patientresponds to a treatment positively when the patient shows reduced or nosymptoms.

As used herein, “Beck Depression Inventory” or “BDI” refers to an indexsystem for assessing the depression or response of a multiple sclerosispatient. The index consists of 21 questions scored from 0-3 so that thetotal index score ranges from 0-63; 1-10: ups and downs are considerednormal; 11-16: mild mood disturbance; 17-20: borderline clinicaldepression; 21-30: moderate depression; 31-40: severe depression; over40: extreme depression.

As used herein, “Multiple Sclerosis Impact Scale” or “MSIS-29” refers toan index system for assessing the symptomatic severity or response of amultiple sclerosis patient. The index assesses the physical andpsychological impact of multiple sclerosis. See Hobert et al., TheMultiple Sclerosis Impact Scale (MSIS-29): a new patient-based outcomemeasure. Brain 2001; 124:962-73. Each variable is scored from 0-5 sothat the total index score ranges from 0-145.

As used herein, “eukaryotic” refers to belonging to a cell that containsa nucleus and membrane-bound organelles.

As used herein, “bacteria,” “bacterium,” and “archaea” refer tosingle-celled prokaryotes that lack membrane bound nuclei and lackorganelles.

As used herein, “colony forming units” (cfu) refers to an estimate ofthe number of viable microorganism cells in a given sample.

As used herein, “viable” means possessing the ability to multiply.

As used herein, “fecal bacteria” refers to bacteria that can be found infecal matter.

As used herein, “isolated” or “purified” refers to a bacterium or otherentity or substance that has been (1) separated from at least some ofthe components with which it was associated when initially produced(whether in nature or in an experimental setting), and/or (2) produced,prepared, purified, and/or manufactured by the hand of man. Isolated orpurified bacteria can be separated from at least about 10%, about 20%,about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about90%, or more of the other components with which they were initiallyassociated.

As used herein, “cytotoxic” activity or bacterium includes the abilityto kill a bacterial cell, such as a pathogenic bacterial cell. A“cytostatic” activity or bacterium includes the ability to inhibit,partially or fully, growth, metabolism, and/or proliferation of abacterial cell, such as a pathogenic bacterial cell.

As used herein, the terms “pathogen” and “pathogenic” in reference to abacterium or any other organism or entity includes any such organism orentity that is capable of causing or affecting a disease, disorder orcondition of a host organism containing the organism or entity.

As used herein, “spore” or a population of “spores” includes bacteria(or other single-celled organisms) that are generally viable, moreresistant to environmental influences such as heat and bacteriocidalagents than vegetative forms of the same bacteria, and typically capableof germination and out-growth. “Spore-formers” or bacteria “capable offorming spores” are those bacteria containing the genes and othernecessary abilities to produce spores under suitable environmentalconditions.

As used herein, a “combination” of two or more bacteria includes thephysical co-existence of the two bacteria, either in the same materialor product or in physically connected products, as well as the temporalco-administration or co-localization of the two bacteria.

As used herein, “subject” refers to any animal subject including humans,laboratory animals (e.g., primates, rats, mice), livestock (e.g., cows,sheep, goats, pigs, turkeys, chickens), and household pets (e.g., dogs,cats, rodents, etc.). The subject or patient may be healthy, or may besuffering from an infection due to a gastrointestinal pathogen or may beat risk of developing or transmitting to others an infection due to agastrointestinal pathogen.

As used herein, “Shannon Diversity Index” refers to a diversity indexthat accounts for abundance and evenness of species present in a givencommunity using the formula

$H = {- {\sum\limits_{i = 1}^{R}{p_{i}\ln \; p_{i}}}}$

where H is Shannon Diversity Index, R is the total number of species inthe community, and p_(i) is the proportion of R made up of the ithspecies. Higher values indicate diverse and equally distributedcommunities, and a value of 0 indicates only one species is present in agiven community. For further reference, see Shannon and Weaver, (1949)The mathematical theory of communication. The University of IllinoisPress, Urbana. 117 pp.

As used herein, “antibiotic” refers to a substance that is used to treatand/or prevent bacterial infection by killing bacteria, inhibiting thegrowth of bacteria, or reducing the viability of bacteria.

As used herein, an “intermittent dosing schedule” means that atherapeutic composition is administered for a period of time followed bya period of time (a treatment period) where treatment with suchtherapeutic composition is withheld (a rest period). Intermittent dosingregimens can be expressed as treatment period in days or weeks/restperiod in days or weeks. For example, a 4/1 intermittent dosing schedulerefers to an intermittent dosing schedule where the treatment period isfour weeks/days and the rest period is one week/day.

As used herein, a “continuous dosing schedule” refers to a dosingschedule where a therapeutic composition is administered during atreatment period without a rest period. Throughout the treatment periodof a continuous dosing schedule, a therapeutic composition can beadministered, for example, daily, or every other day, or every thirdday. On a day when a therapeutic composition is administered, it can beadministered in a single dose, or in multiple doses throughout the day.

As used herein, “dosing frequency” refers to the frequency ofadministering doses of a therapeutic composition in a given time. Dosingfrequency can be indicated as the number of doses per a given time, forexample, once per day, once a week, or once in two weeks.

As used herein, “dosing interval” refers to the amount of time thatelapses between multiple doses being administered to a subject.

Multiple sclerosis is a pro-inflammatory demyelinating disease of thecentral nervous system. Damage to the myelin forms scar tissue whichlead to distorted or interrupted nerve impulses traveling to and fromthe brain and spinal cord, producing a wide variety of symptoms.

Multiple sclerosis symptoms include bladder dysfunction (e.g.,difficulty urinating, frequent urination, urine leakage), bladderinfection, bowel dysfunction (e.g., constipation and stool leakage),depression, dizziness, vertigo, fatigue, itching, pain, sexualdiscomfort, tremors, walking difficulties, muscle spasms, brain lesions,double vision, eye discomfort, rapid eye movements, vision loss, memoryloss, difficulty solving problems, hearing loss, slurred speech,difficulty with chewing or difficulty with swallowing. Multiple symptomsmay co-exist in the same patient.

Multiple sclerosis occurs most often in people ages 20 to 40, althoughthe disease may afflict people of any age. It affects women morefrequently than men and appears to run in some families.

Different types of multiple sclerosis exist. As used herein, “clinicallyisolated syndrome” refers to a disease form characterized by the firstepisode of neurologic symptoms caused by inflammation and damage tomyelin. The episode, which must last for at least 24 hours, ischaracteristic of multiple sclerosis but fails to meet the diagnosiscriteria of multiple sclerosis. The subject may or may not subsequentlydevelop multiple sclerosis. Subjects with clinically isolated syndromecan present with lesions on the brain which increase the chances of thesubject having a subsequent episode and developing multiple sclerosis.

As used herein, “relapsing-remitting multiple sclerosis” refers to themost common disease type, characterized by defined episodes involvingnew or more advanced neurologic symptoms. During periods in betweenepisodes subjects show signs of partial or complete remission.Eighty-five percent (85%) of patients present with relapsing-remittingmultiple sclerosis.

As used herein, “primary progressive multiple sclerosis” refers to asubject with continuous worsening neurologic function overtime withremissions or obvious relapses. There may be temporary plateaus duringwhich the disease does not progress. Patients are identified as activeat the time of relapses or new MRI lesions or not active. Primaryprogressive multiple sclerosis includes patients who were previouslydiagnosed with progressive-relapsing multiple sclerosis.

As used herein, “secondary progressive multiple sclerosis” refers to asubject with worsening neurologic function overtime. Secondaryprogressive multiple sclerosis follows an initial relapsing-remittingcourse.

Several theories have been proposed for the cause of multiple sclerosis.There is some evidence to suggest that the body's immune system reactsto an environmental or infectious agent in genetically susceptibleindividuals causing inflammation and demyelination in the centralnervous system.

The most common symptoms of multiple sclerosis are bladder dysfunction,bladder infection, bowel dysfunction, depression, dizziness, vertigo,fatigue, itching, pain, sexual discomfort, tremors, walkingdifficulties, and muscle spasms. These symptoms occur in between periodswhen the symptoms go away (remissions) and are referred to as“flare-ups”, “relapse”, or “exacerbation”. A relapse can last at least24 hours and are separated from the previous relapse by at least 30days. Signs of a relapse can include a subject with multiple sclerosisexperiencing fatigue, tingling, brain fog, muscle spasms, or depression.A relapse can last for days, weeks, or months, followed by periods ofremission.

People suffering from acute multiple sclerosis attacks may be treatedwith corticosteroids. Prednisone, methylprednisolone, budesonide, andhydrocortisone are corticosteroids used to reduce inflammation. They canbe given orally, intravenously, through an enema, or in a suppository,depending on the location of the inflammation. Corticosteroids can causeside effects such as weight gain, acne, facial hair, hypertension,diabetes, mood swings, and increased risk of infection, so doctorscarefully monitor patients taking these medications.

Multiple sclerosis symptoms can be severe enough that the person must behospitalized. In fact, after the first hospitalization for diagnosis ofMS, patients usually experience hospitalization several times due torepeated attacks of the disease. The patient may need a special diet,feeding through a vein, medications, or sometimes surgery.

Recently, relapsing-remitting multiple sclerosis patients wereidentified as having a distinct gut microbiota compared to healthypatients as described in Chen, et al., Multiple Sclerosis patients havea distinct gut microbiota compared to healthy controls, ScientificReports 2016; (6): 1-10. Many chronic diseases and disorders of the GItract have chronic infection/infestation as their underlyingpathological conditions (e.g., multiple sclerosis). In one aspect, thepresent disclosure includes and relates to the use of a fecalmicrobiota, one or more microbial species therefrom, an active fragmentor component therefrom for the treatment and/or prophylaxis of variousdisease states (e.g., multiple sclerosis) related to the presence of‘abnormal’ microflora in the GI tract. An active fragment of a bacteriumcan be any active molecule isolated from such bacteria by any knownmethod for preparing/identifying active fragments of bacteria andproteins secreted from bacteria. Such methods include but are notlimited to the following: sonication, osmotic shock, detergent lysis,high pressure, transfer appropriate DNA to other organisms, such asbacteria, plant or animal that is then used as a feed additive asdescribed previously. In one aspect, an active fragment or component ofa bacterium is selected from the group consisting of a mycolate or aderivative thereof, a polysaccharide, a lipoglycan, a small peptide, athiopeptide, a protein, a nucleic acid molecule, a metabolite, a cellwall component, or any combination thereof. In one aspect, an activefragment is a protein or a secretion. In another aspect, an activefragment is a secreted protein.

In one aspect, the present disclosure provides a method for treatingmultiple sclerosis in a subject in need thereof, where the methodcomprises administering to the subject a pharmaceutically active dose ofa therapeutic composition comprising live non-pathogenic fecal bacteria.In another aspect, this disclosure provides use of a compositioncomprising live non-pathogenic fecal bacteria in the manufacture of amedication for the treatment of multiple sclerosis. In one aspect, amethod is for treating a form of multiple sclerosis selected from thegroup consisting of clinically isolated syndrome (CIS),relapsing-remitting MS (RRMS), primary progressive MS (PPMS), andsecondary progressive MS (SPMS). In one aspect, a therapeuticcomposition comprises an isolated or purified population of livenon-pathogenic fecal bacteria. In one aspect, a therapeutic compositioncomprises a non-selected fecal microbiota. In another aspect, atherapeutic composition comprises a non-selected and substantiallycomplete fecal microbiota. In another aspect, a therapeutic compositioncomprises a full-spectrum fecal microbiota. In one aspect, a methodfurther comprises administering, onabotulinumtoxin A, desmopressin,tolterodine, oxybutynin, darifenacin, tamsulosin, terazosin, prazosin,oxybutynin, propantheline, trospium chloride, imipramine, solifenacinsuccinate, sulfamethoxazole, ciprofloxacin, nitrofurantoin, methenamine,phenazopyridine, docusate, bisacodyl, docusate stool softener laxative,sodium phosphate, Mineral Oil, psyllium hydrophilic musilloid, magnesiumhydroxide, glycerin, duloxetine hydrochloride, venlafaxine, paroxetine,bupropion, sertraline, meclizine, dextromethorphan quinidine, modafinil,fluoxetine, amantadine, hydroxyzine, phenytoin, amitriptyline,clonazepam, gabapentin, nortriptyline, carbamazepine, tadalafil,vardenafil, papaverine, alprostadil, sildenafil, dantrolene, baclofen,diazepam, tizanidine, isoniazid, dalfampridine, and a combinationthereof. In another aspect, a method further comprises administering oneor more injectable medications selected from the group consisting ofinterferon beta-la, interferon beta-1b, glatiramer acetate,peginterferon beta-1a, daclizumab, and immune globulin. In anotheraspect, a method further comprises administering one or more oralmedications selected from the group consisting of teriflunomide,fingolimod, and dimethyl fumarate. In a further aspect, a method furthercomprises administering one or more infused medications selected fromthe group consisting of alemtuzumab, mitoxantrone, and natalizumab.

In one aspect, the present disclosure provides a method which eliminatesor reduces one or more multiple sclerosis symptoms selected from thegroup consisting of bladder dysfunction, bladder infection, boweldysfunction, dizziness, vertigo, fatigue, itching, pain, sexualdiscomfort, tremors, walking difficulties, muscle spasms, and acombination thereof. In another aspect, the present disclosure providesa method which eliminates or reduces one or more multiple sclerosissymptoms selected from the group consisting of constipation and stoolleakage, difficulty urinating, frequent urination, urine leakage, doublevision, eye discomfort, rapid eye movements, vision loss, memory loss,difficulty solving problems, depression, hearing loss, slurred speech,difficulty with chewing and difficulty with swallowing.

In another aspect, a method further comprises administering medicationselected from the group consisting of onabotulinumtoxin A, desmopressin,tolterodine, oxybutynin, darifenacin, tamsulosin, terazosin, prazosin,oxybutynin, propantheline, trospium chloride, imipramine, solifenacinsuccinate and a combination thereof. In another aspect, a method furthercomprises administering medication selected from the group consisting ofsulfamethoxazole, ciprofloxacin, nitrofurantoin, methenamine,phenazopyridine, and a combination thereof. In another aspect, a methodfurther comprises administering medication selected from the groupconsisting of docusate, bisacodyl, docusate stool softener laxative,sodium phosphate, Mineral Oil, psyllium hydrophilic musilloid, magnesiumhydroxide, glycerin, and a combination thereof. In another aspect, amethod further comprises administering medication selected from thegroup consisting of duloxetine hydrochloride, venlafaxine, paroxetine,fluoxetine, bupropion, and sertraline. In another aspect, a methodfurther comprises administering medication selected from the groupconsisting of meclizine and dextromethorphan quinidine. In a furtheraspect, a method further comprises administering medication selectedfrom the group consisting of modafinil, fluoxetine, and amantadine. Inanother aspect, a method further comprises administering hydroxyzine. Inanother aspect, a method further comprises administering medicationselected from the group consisting of phenytoin, amitriptyline,clonazepam, gabapentin, nortriptyline, and carbamazepine. In anotheraspect, a method further comprises administering medication selectedfrom the group consisting of tadalafil, vardenafil, papaverine,alprostadil, and sildenafil. In another aspect, a method furthercomprises administering medication selected from the group consisting ofonabotulinumtoxin A, dantrolene, baclofen, clonazepam, baclofen,diazepam, and tizanidine. In a further aspect, a method furthercomprises administering medication selected from the group consisting ofisoniazid and clonazepam. In another aspect, a method further comprisesadministering dalfampridine.

In one aspect, the present disclosure provides a method for treatingmultiple sclerosis in a subject in need thereof, where the methodcomprises administering to the subject a pharmaceutically active dose ofa therapeutic composition comprising live non-pathogenic bacteria. Inone aspect, the present disclosure provides a method for treatingmultiple sclerosis in a subject in need thereof, where the methodcomprises administering daily to the subject a pharmaceutically activedose of a therapeutic composition comprising live non-pathogenic fecalbacteria. In one aspect, a therapeutic composition is administered to amultiple sclerosis patient in need thereof at least once daily or weeklyfor at least two consecutive days or weeks. In one aspect, a therapeuticcomposition is administered at least once daily or weekly for at least3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 consecutive days orweeks. In another aspect, a therapeutic composition is administered atleast once daily or weekly for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, or 12 consecutive days or weeks. In one aspect, a therapeuticcomposition is administered at least once daily or weekly for at most 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 consecutivedays or weeks. In another aspect, a therapeutic composition isadministered at least once daily or weekly for at most 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, or 12 consecutive weeks or months. In a further aspect,a therapeutic composition is administered at least once for at least 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 consecutive months or years,chronically for a subject's entire life span, or an indefinite period oftime.

In one aspect, a therapeutic composition is administered to a multiplesclerosis patient in need thereof at least twice daily or weekly for atleast two consecutive days or weeks. In one aspect, a therapeuticcomposition is administered at least twice daily or weekly for at least3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 consecutive days orweeks. In another aspect, a therapeutic composition is administered atleast twice daily or weekly for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, or 12 consecutive days or weeks. In one aspect, a therapeuticcomposition is administered at least twice daily or weekly for at most4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20consecutive days or week. In another aspect, a therapeutic compositionis administered at least twice daily or weekly for at most 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, or 12 consecutive weeks or months. In a furtheraspect, a therapeutic composition is administered at least twice for atleast 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 consecutive months oryears, chronically for a subject's entire life span, or an indefiniteperiod of time.

In one aspect, a therapeutic composition is administered to a multiplesclerosis patient in need thereof at least three times daily or weeklyfor at least two consecutive days or weeks. In one aspect, a therapeuticcomposition is administered at least three times daily or weekly for atleast 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 consecutive days orweeks. In another aspect, a therapeutic composition is administered atleast three times daily or weekly for at least 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, or 12 consecutive days or weeks. In one aspect, a therapeuticcomposition is administered at least three times daily for at most 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 consecutivedays or weeks. In another aspect, a therapeutic composition isadministered at least three times daily for at most 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, or 12 consecutive weeks or months. In a further aspect, atherapeutic composition is administered at least three times for atleast 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 consecutive months oryears, chronically for a subject's entire life span, or an indefiniteperiod of time.

In one aspect, the present disclosure provides a method for treatingmultiple sclerosis in a subject in need thereof, where the methodcomprises administering orally to the subject a pharmaceutically activedose of a therapeutic composition comprising live, non-pathogenic,synthetic bacterial mixture or live, non-pathogenic, purified orextracted, fecal microbiota, where the dose is administered at a dosingschedule of at least once or twice daily or weekly for at least threeconsecutive days or weeks. In another aspect, a dose is administered atleast once, twice, or three times daily or weekly for a period between 1and 12 weeks, between 2 and 12 weeks, between 3 and 12 weeks, between 4and 12 weeks, between 5 and 12 weeks, between 6 and 12 weeks, between 7and 12 weeks, between 8 and 12 weeks, between 9 and 12 weeks, between 10and 12 weeks, between 1 and 2 weeks, between 2 and 3 weeks, between 3and 4 weeks, between 4 and 5 weeks, between 5 and 6 weeks, between 6 and7 weeks, between 7 and 8 weeks, between 8 and 9 weeks, between 9 and 10weeks, or between 10 and 11 weeks.

In one aspect, the present disclosure provides a method for treatingmultiple sclerosis in a subject in need thereof, where the methodcomprises a first dosing schedule followed by a second dosing schedule.In one aspect, a first dosing schedule comprises a treatment orinduction dose. In one aspect, a first dosing schedule comprises acontinuous dosing schedule. In another aspect, a second dosing schedulecomprises a maintenance dose lower than or equal to a pharmaceuticallyactive dose of a first dosing schedule. In another aspect, a seconddosing schedule lasts for at least about 2, 4, 6, 8, 10, 12, 18, 24, 36,48, 72, or 96 months. In one aspect, a second dosing schedule lastspermanently, for a treated subject's entire life span, or an indefiniteperiod of time. In one aspect, a second dosing schedule is a continuousdosing schedule. In another aspect, a second dosing schedule is anintermittent dosing schedule. In a further aspect, a second dosingschedule is an intermittent dosing schedule comprising a treatmentperiod of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 daysfollowed by a resting period of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, or 14 days. In another aspect, a second dosing schedulecomprises administering a second dose (e.g., a maintenance dose) everyother day, every two days, or every 3, 4, 5, 6, 7, 8 days. In anotheraspect, a maintenance dose is administered for an extended period oftime with or without titration (or otherwise changing the dosage ordosing schedule). In one aspect, the interval between a first and asecond dosing schedule is at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, or 12 weeks. In another aspect, a second dosing schedule (e.g., amaintenance dose) comprises a dosage about 2, 5, 10, 50, 100, 200, 400,800, 1000, 5000 or more folds lower than the dosage used in a firstdosing schedule (e.g., an initial treatment dose). In another aspect, asecond dosing schedule (e.g., a maintenance dosing schedule) has anequal or lower dosing frequency than a first dosing schedule (e.g., aninitial treatment dosing schedule). In another aspect, a second dosingschedule (e.g., a maintenance dosing schedule) has a higher dosinginterval than a first dosing schedule (e.g., an initial treatment dosingschedule).

In one aspect, a first or second dosing schedule used in a method can beonce-a-week, twice-a-week, or thrice-a-week. The term “once-a-week”means that a dose is administered once in a week, preferably on the sameday of each week. “Twice-a-week” means that a dose is administered twotimes in a week, preferably on the same two days of each weekly period.“Thrice-a-week” means that a dose is administered three times in a week,preferably on the same three days of each weekly period.

In one aspect, a subject being treated is a subject already withmultiple sclerosis. Administration of a disclosed therapeuticcomposition to clinically, asymptomatic human subject who is geneticallypredisposed or prone to multiple sclerosis is also useful in preventingthe onset of clinical symptoms of multiple sclerosis. A human subjectgenetically predisposed or prone to multiple sclerosis can be a humansubject having a close family member or relative exhibiting or havingsuffered multiple sclerosis. In another aspect, a subject being treatedis a subject in which multiple sclerosis is to be prevented. In anotheraspect, a subject being treated is predisposed or susceptible tomultiples sclerosis. In another aspect, a subject being treated is asubject diagnosed as having multiple sclerosis. In one aspect, a subjectbeing treated is a patient in need thereof. In another aspect, a patientbeing treated is immunocompromised.

In one aspect, a subject being treated is a human patient. In oneaspect, a patient is a male patient. In one aspect, a patient is afemale patient. In one aspect, a patient is a premature newborn. In oneaspect, a patient is a term newborn. In one aspect, a patient is aneonate. In one aspect, a patient is an infant. In one aspect, a patientis a toddler. In one aspect, a patient is a young child. In one aspect,a patient is a child. In one aspect, a patient is an adolescent.

In one aspect, a patient is a pediatric patient. In one aspect, apatient is a geriatric patient. In one aspect, a human patient is achild patient below about 18, 15, 12, 10, 8, 6, 4, 3, 2, or 1 year old.In another aspect, a human patient is an adult patient. In anotheraspect, a human patient is an elderly patient. In a further aspect, ahuman patient is a patient above about 30, 35, 40, 45, 50, 55, 60, 65,70, 75, 80, 85, 90, or 95 years old. In another aspect, a patient isabout between 1 and 5, between 2 and 10, between 3 and 18, between 21and 50, between 21 and 40, between 21 and 30, between 50 and 90, between60 and 90, between 70 and 90, between 60 and 80, or between 65 and 75years old. In one aspect, a patient is a young old patient (65-74years). In one aspect, a patient is a middle old patient (75-84 years).In one aspect, a patient is an old old patient (>85 years).

In one aspect, a method comprises administering a therapeuticcomposition orally, by enema, or via rectal suppository. In one aspect,a therapeutic composition administered herein is formulated as anenteric coated (and/or acid-resistant) capsule or microcapsule, orformulated as part of or administered together with a food, a foodadditive, a dairy-based product, a soy-based product or a derivativethereof, a jelly, flavored liquid, ice block, ice cream, or a yogurt. Inanother aspect, a therapeutic composition administered herein isformulated as an acid-resistant enteric coated capsule. A therapeuticcomposition can be provided as a powder for sale in combination with afood or drink. A food or drink can be a dairy-based product or asoy-based product. In another aspect, a food or food supplement containsenteric-coated and/or acid-resistant microcapsules containing atherapeutic composition.

In an aspect, a therapeutic composition comprises a liquid culture. Inanother aspect, a therapeutic composition is homogenized, lyophilized,pulverized and powdered. It may then be infused, dissolved such as insaline, as an enema. Alternatively the powder may be encapsulated asenteric-coated and/or acid-resistant delayed release capsules for oraladministration. In an aspect, the powder may be double encapsulated withacid-resistant/delayed release capsules for oral administration. Thesecapsules may take the form of enteric-coated and/or acid-resistantdelayed release microcapsules. A powder can preferably be provided in apalatable form for reconstitution for drinking or for reconstitution asa food additive. In a further aspect, a food is yogurt. In one aspect, apowder may be reconstituted to be infused via naso-duodenal infusion.

In another aspect, a therapeutic composition administered herein is in aliquid, frozen, freeze-dried, spray-dried, foam-dried, lyophilized, orpowder form. In a further aspect, a therapeutic composition administeredherein is formulated as a delayed or gradual enteric release form. Inanother aspect, a therapeutic composition administered herein comprisesan excipient, a saline, a buffer, a buffering agent, or afluid-glucose-cellobiose agar (RGCA) media. In another aspect, atherapeutic composition administered herein comprises a cryoprotectant.In one aspect, a cryoprotectant comprises polyethylene glycol, skimmilk, erythritol, arabitol, sorbitol, glucose, fructose, alanine,glycine, proline, sucrose, lactose, ribose, trehalose, dimethylsulfoxide (DMSO), glycerol, or a combination thereof.

In one aspect, a therapeutic composition administered herein furthercomprises an acid suppressant, an antacid, an H2 antagonist, a protonpump inhibitor or a combination thereof. In one aspect, a therapeuticcomposition administered herein substantially free of non-living matter.In another aspect, a therapeutic composition administered hereinsubstantially free of acellular material selected from the groupconsisting of residual fiber, DNA, viral coat material, and non-viablematerial.

In one aspect, a therapeutic composition also comprises or issupplemented with a prebiotic nutrient selected from the groupconsisting of polyols, fructooligosaccharides (FOSs), oligofructoses,inulins, galactooligosaccharides (GOSs), xylooligosaccharides (XOSs),polydextroses, monosaccharides, tagatose, and/or mannooligosaccharides.

In one aspect, a method further comprises pretreating a subject with anantibiotic composition prior to administering a therapeutic bacterial ormicrobiota composition. In one aspect, an antibiotic compositionadministered herein comprises an antibiotic selected from the groupconsisting of rifabutin, clarithromycin, clofazimine, vancomycin,rifampicin, nitroimidazole, chloramphenicol, and a combination thereof.In another aspect, an antibiotic composition administered hereincomprises an antibiotic selected from the group consisting of rifaximin,rifamycin derivative, rifampicin, rifabutin, rifapentine, rifalazil,bicozamycin, aminoglycoside, gentamycin, neomycin, streptomycin,paromomycin, verdamicin, mutamicin, sisomicin, netilmicin, retymicin,kanamycin, aztreonam, aztreonam macrolide, clarithromycin,dirithromycin, roxithromycin, telithromycin, azithromycin, bismuthsubsalicylate, vancomycin, streptomycin, fidaxomicin, amikacin,arbekacin, neomycin, netilmicin, paromomycin, rhodostreptomycin,tobramycin, apramycin, and a combination thereof. In a further aspect, amethod further comprises pretreating a subject with an anti-inflammatorydrug prior to administration of a therapeutic bacterial or microbiotacomposition.

In one aspect, a method achieves a remission, cure, response, orresolution rate of multiple sclerosis of at least about 10%, 15%, 20%,25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%,95%, 97%, or 99%. In one aspect, a treatment method achieves at least10%, 20%, 30%, 50%, 60%, 70%, 80%, or 90% reduction of multiplesclerosis disease symptoms after 4, 8, or 12 weeks of treatment comparedto baseline (e.g., immediately prior to treatment). In one aspect, atreatment method achieves at least 10%, 20%, 30%, 50%, 60%, 70%, 80%, or90% reduction of multiple sclerosis disease symptoms in at least 10%,20%, 30%, 50%, 60%, 70%, 80%, or 90% patients after 4, 8, or 12 weeks oftreatment compared to baseline (e.g., immediately prior to treatment).In an aspect, the present disclosure provides a method which achieves a10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% reduction of a patient'sBDI score after 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks or monthsof treatment. In another aspect, a method achieves a 10-20%, 20-30%,30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-99% reduction of apatient's BDI score after 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeksor months of treatment. In an aspect, the present disclosure provides amethod which achieves a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%reduction of a patient's MSIS-29 score after 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, or 12 weeks or months of treatment. In another aspect, a methodachieves a 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%,80-90%, or 90-99% reduction of a patient's MSIS-29 score after 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks or months of treatment.

In one aspect, a method achieves a remission, cure, response, orresolution rate of multiple sclerosis of between about 10-20%, 20-30%,30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-99%. In oneaspect, a treatment method achieves between about 10-20%, 20-30%,30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-99% reduction ofmultiple sclerosis disease symptoms after 4, 8, or 12 weeks of treatmentcompared to baseline (e.g., immediately prior to treatment). In oneaspect, a treatment method achieves between about 10-20%, 20-30%,30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-99% reduction ofmultiple sclerosis disease symptoms in about 10-20%, 20-30%, 30-40%,40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-99% of patients after 4,8, or 12 weeks of treatment compared to baseline (e.g., immediatelyprior to treatment).

In a further aspect, a patient is assessed using the Beck DepressionInventory (BDI) or the Multiple Sclerosis Impact Scale (MSIS-29) asdescribed in Hobert et al., The Multiple Sclerosis Impact Scale(MSIS-29): a new patient-based outcome measure. Brain 2001;124:962-73.In one aspect, a treatment method achieves at least 10%, 20%, 30%, 50%,60%, 70%, 80%, or 90% reduction of Beck Depression Inventory Score after4, 8, or 12 weeks of treatment compared to baseline (e.g., immediatelyprior to treatment). In one aspect, a treatment method achieves at least10%, 20%, 30%, 50%, 60%, 70%, 80%, or 90% reduction of Beck DepressionInventory score in at least 10%, 20%, 30%, 50%, 60%, 70%, 80%, or 90%patients after 4, 8, or 12 weeks of treatment compared to baseline(e.g., immediately prior to treatment). In one aspect, a treatmentmethod achieves at least 10%, 20%, 30%, 50%, 60%, 70%, 80%, or 90%reduction of MSIS-29 Score after 4, 8, or 12 weeks of treatment comparedto baseline (e.g., immediately prior to treatment). In one aspect, atreatment method achieves at least 10%, 20%, 30%, 50%, 60%, 70%, 80%, or90% reduction of MSIS-29 score in at least 10%, 20%, 30%, 50%, 60%, 70%,80%, or 90% patients after 4, 8, or 12 weeks of treatment compared tobaseline (e.g., immediately prior to treatment).

In one aspect, a treatment method achieves between about 10-20%, 20-30%,30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-99% reduction ofBeck Depression Inventory Score after 4, 8, or 12 weeks of treatmentcompared to baseline (e.g., immediately prior to treatment). In oneaspect, a treatment method achieves between about 10-20%, 20-30%,30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-99% reduction ofBeck Depression Inventory score in between about 10-20%, 20-30%, 30-40%,40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-99% of patients after 4,8, or 12 weeks of treatment compared to baseline (e.g., immediatelyprior to treatment). In one aspect, a treatment method achieves betweenabout 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or90-99% reduction of MSIS-29 Score after 4, 8, 12 weeks of treatmentcompared to baseline (e.g., immediately prior to treatment). In oneaspect, a treatment method achieves between about 10-20%, 20-30%,30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-99% reduction ofMSIS-29 score in between about 10-20%, 20-30%, 30-40%, 40-50%, 50-60%,60-70%, 70-80%, 80-90%, or 90-99% of patients after 4, 8, or 12 weeks oftreatment compared to baseline (e.g., immediately prior to treatment).

In one aspect, every about 200mg of a pharmaceutical compositioncomprises a pharmacologically active dose. In one aspect, every about75, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, 750, 1000,1500, or 2000 mg of a pharmaceutical composition comprises apharmacologically active dose.

In one aspect, a pharmaceutically active or therapeutic effective dosecomprises at least about 10⁵, 10⁶, 10⁷, 10⁸, 10⁹, 10¹⁰, 10¹¹, 10¹²,10¹³, 10¹⁴, or 10¹⁵ cfu. In another aspect, a pharmaceutically activetherapeutic effective dose comprises at most about 10⁵, 10⁶, 10⁷, 10⁸,10⁹, 10¹⁰, 10¹¹, 10¹², 10¹³, 10¹⁴, or 10¹⁵ cfu. In a further aspect, apharmacologically active therapeutic effective dose is selected from thegroup consisting of from 10⁸ cfu to 10¹⁴ cfu, from 10⁹ cfu to 10¹³ cfu,from 10¹⁰ cfu to 10¹² cfu, from 10⁹ cfu to 10¹⁴ cfu, from 10⁹ cfu to10¹² cfu, from 10⁹ cfu to 10¹¹ cfu, from 10⁹ cfu to 10¹⁰ cfu, from 10¹⁰cfu to 10¹⁴ cfu, from 10¹⁰ cfu to 10¹³ cfu, from 10¹¹ cfu to 10¹⁴ cfu,from 10¹¹ cfu to 10¹³ cfu, from 10¹² cfu to 10¹⁴ cfu, and from 10¹³ cfuto 10¹⁴ cfu. In one aspect, a pharmaceutical composition comprises theforegoing pharmaceutically active or therapeutic effective dose in aunit weight of about 0.2, 0.4, 0.6, 0.8 or 1.0 gram, or a unit volume ofabout 0.2, 0.4, 0.6, 0.8 or 1.0 milliliter.

In one aspect, a pharmaceutically active or therapeutic effective dosecomprises at least about 10⁵, 10⁶, 10⁷, 10⁸, 10⁹, 10¹⁰, 10¹¹, 10¹²,10¹³, 10¹⁴, or 10¹⁵ cells or spores. In another aspect, apharmaceutically active or therapeutic effective dose comprises at mostabout 10⁵, 10⁶, 10⁷, 10⁸, 10⁹, 10¹⁰, 10¹¹, 10¹², 10¹³, 10¹⁴, or 10¹⁵total cells or spores. In a further aspect, a pharmacologically activeor therapeutic effective dose is selected from the group consisting offrom 10⁸ to 10¹⁴, from 10⁹ to 10¹³, from 10¹⁰ to 10¹², from 10⁹ to 10¹⁴,from 10⁹ to 10¹², from 10⁹ to 10¹¹, from 10⁹ to 10¹⁰, from 10¹⁰ to 10¹⁴,from 10¹⁰ to 10¹³, from 10¹¹ to 10¹⁴, from 10¹¹ to 10¹³, from 10¹² to10¹⁴, and from 10¹³ to 10¹⁴ cells or spores. In an aspect, thepharmaceutically active or therapeutic effective dose cell count isdirected to live cells. In one aspect, a pharmaceutical compositioncomprises the foregoing pharmaceutically active or therapeutic effectivedose in a unit weight of about 0.2, 0.4, 0.6, 0.8 or 1.0 gram, or a unitvolume of about 0.2, 0.4, 0.6, 0.8 or 1.0 milliliter. In an aspect, apharmaceutically active or therapeutic effective dose comprises between10¹⁰ and 10¹² cells. In another aspect, a pharmaceutically active ortherapeutic effective dose comprises between 10¹⁰ and 10¹² cells percapsule. In another aspect, a pharmaceutically active or therapeuticeffective dose comprises between 10¹¹ and 10¹² cells per capsule. In afurther aspect, a pharmaceutically active or therapeutic effective dosecomprises between 10⁹ and 10¹² cells per capsule.

In one aspect, a therapeutic composition administered herein comprisesfecal bacteria. In one aspect, a therapeutic composition administeredherein comprises one or more, two or more, three or more, four or more,or five or more isolated, purified, or cultured microorganisms selectedfrom the group consisting of Clostridium, Bacillus, Collinsella,Bacteroides, Eubacterium, Fusobacterium, Propionibacterium,Lactobacillus, Ruminococcus, Escherichia coli, Gemmiger, Desulfomonas,Peptostreptococcus, Bifidobacterium, Coprococcus, Dorea, and Monilia.

In one aspect, a therapeutic composition administered herein comprisesat least one, at least two, at least three, at least four, at leastfive, at least six, or at least seven fecal microorganisms selected fromthe group consisting of a Bacteroides fragilis ssp. vulgatus,Collinsella aerofaciens, Bacteroides fragilis ssp. thetaiotaomicron,Peptostreptococcus productus II, Parabacteroides distasonis,Fusobacterium prausnitzii, Coprococcus eutactus, Collinsella aerofaciensIII, Peptostreptococcus productus I, Ruminococcus bromii,Bifidobacterium adolescentis, Gemmiger formicilis, Bifidobacteriumlongum, Eubacterium siraeum, Ruminococcus torques, Eubacterium rectale,Eubacterium eligens, Bacteroides eggerthii, Clostridium leptum,Bacteroides fragilis ssp. A, Eubacterium biforme, Bifidobacteriuminfantis, Eubacterium rectale Coprococcus comes, Pseudoflavonifractorcapillosus, Ruminococcus albus, Dorea formicigenerans, Eubacteriumhallii, Eubacterium ventriosum I, Fusobacterium russi, Ruminococcusobeum, Eubacterium rectale, Clostridium ramosum, Lactobacillusleichmannii, Ruminococcus callidus, Butyrivibrio crossotus,Acidaminococcus fermentans, Eubacterium ventriosum, Bacteroides fragilisssp. fragilis, Bacteroides AR, Coprococcus catus, Aerostipes hadrus,Eubacterium cylindroides, Eubacterium ruminantium, Eubacterium CH-1,Staphylococcus epidermidis, Peptostreptococcus BL, Eubacterium limosum,Tissirella praeacuta, Bacteroides L, Fusobacterium mortiferum I,Fusobacterium naviforme, Clostridium innocuum, Clostridium ramosum,Propionibacterium acnes, Ruminococcus flavefaciens, Ruminococcus AT,Peptococcus AU-1, Bacteroides fragilis ssp. ovatus, -ssp. d, -ssp. f;Bacteroides L-1, L-5; Fusobacterium nucleatum, Fusobacterium mortiferum,Escherichia coli, Gemella morbillorum, Finegoldia magnus, Peptococcus G,-AU-2; Streptococcus intermedius, Ruminococcus lactaris, Ruminococcus COGemmiger X, Coprococcus BH, -CC; Eubacterium tenue, Eubacterium ramulus,Bacteroides clostridiiformis ssp. clostridliformis, Bacteroidescoagulans, Prevotella oralis, Prevotella ruminicola, Odoribactersplanchnicus, Desuifomonas pigra, Lactobacillus G, Succinivibrio A, anda combination thereof.

In one aspect, a therapeutic composition administered herein comprisesno viable Bacteroides, Fusobacterium, Propionibacterium, Lactobacillus,Ruminococcus, Escherichia coli, Gemmiger, Desulfomonas,Peptostreptococcus, Bifidobacterium, Monilia, or any combinationthereof. In another aspect, a therapeutic composition administeredherein comprises no viable Bacteroides fragilis ssp. vulgatus,Collinsella aerofaciens, Bacteroides fragilis ssp. thetaiotaomicron,Peptostreptococcus productus II, Parabacteroides distasonis,Fusobacterium prausnitzii, Coprococcus eutactus, Collinsella aerofaciensIII, Peptostreptococcus productus I, Ruminococcus bromii,Bifidobacterium adolescentis, Gemmiger formicilis, Bifidobacteriumlongum, Eubacterium siraeum, Ruminococcus torques, Eubacterium rectale,Eubacterium eligens, Bacteroides eggerthii, Clostridium leptum,Bacteroides fragilis ssp. A, Eubacterium biforme, Bifidobacteriuminfantis, Eubacterium rectale Coprococcus comes, Pseudoflavonifractorcapillosus, Ruminococcus albus, Dorea formicigenerans, Eubacteriumhallii, Eubacterium ventriosum I, Fusobacterium russi, Ruminococcusobeum, Eubacterium rectale, Clostridium ramosum, Lactobacillusleichmannii, Ruminococcus callidus, Butyrivibrio crossotus,Acidaminococcus fermentans, Eubacterium ventriosum, Bacteroides fragilisssp. fragilis, Bacteroides AR, Coprococcus catus, Aerostipes hadrus,Eubacterium cylindroides, Eubacterium ruminantium, Eubacterium CH-1,Staphylococcus epidermidis, Peptostreptococcus BL, Eubacterium limosum,Tissirella praeacuta, Bacteroides L, Fusobacterium mortiferum I,Fusobacterium naviforme, Clostridium innocuum, Clostridium ramosum,Propionibacterium acnes, Ruminococcus flavefaciens, Ruminococcus AT,Peptococcus AU-1, Bacteroides fragilis ssp. ovatus, -ssp. d, -ssp. f;Bacteroides L-1, L-5; Fusobacterium nucleatum, Fusobacterium mortiferum,Escherichia coli, Gemella morbillorum, Finegoldia magnus, Peptococcus G,-AU-2; Streptococcus intermedius, Ruminococcus lactaris, Ruminococcus COGemmiger X, Coprococcus BH, -CC; Eubacterium tenue, Eubacterium ramulus,Bacteroides clostridiiformis ssp. clostridliformis, Bacteroidescoagulans, Prevotella oralis, Prevotella ruminicola, Odoribactersplanchnicus, Desuifomonas pigra, Lactobacillus G, Succinivibrio A, or acombination thereof.

In one aspect, a therapeutic composition administered herein comprises afecal microbiota. In another aspect, the preparation of a fecalmicrobiota used herein involves a treatment selected from the groupconsisting of ethanol treatment, detergent treatment, heat treatment,irradiation, and sonication. In another aspect, the preparation of afecal microbiota used herein involves no treatment selected from thegroup consisting of ethanol treatment, detergent treatment, heattreatment, irradiation, and sonication. In one aspect, the preparationof a fecal microbiota used herein involves a separation step selectedfrom the group consisting of density gradients, filtration (e.g.,sieves, nylon mesh), and chromatography. In another aspect, thepreparation of a fecal microbiota used herein involves no separationstep selected from the group consisting of density gradients, filtration(e.g., sieves, nylon mesh), and chromatography. In another aspect, afecal microbiota used herein comprises a donor's entire fecalmicrobiota. In another aspect, a therapeutic composition administeredherein comprises a fecal microbiota substantially free of eukaryoticcells from the fecal microbiota's donor.

In another aspect, a therapeutic composition administered hereincomprises a fecal microbiota further supplemented, spiked, or enhancedwith a fecal microorganism. In one aspect, a fecal microbiota issupplemented with a non-pathogenic (or with attenuated pathogenicity)bacterium of Clostridium, Collinsella, Dorea, Ruminococcus, Coprococcus,Prevotella, Veillonella, Bacteroides, Baccillus, or a combinationthereof. In another aspect, a therapeutic composition administeredherein comprises a fecal microbiota further supplemented, spiked, orenhanced with a species of Veillonellaceae, Firmicutes,Gammaproteobacteria, Bacteroidetes, or a combination thereof. In anotheraspect, a therapeutic composition administered herein comprises a fecalmicrobiota further supplemented with fecal bacterial spores. In oneaspect, fecal bacterial spores are Clostridium spores, Bacillus spores,or both.

In an aspect, a therapeutic composition comprises a fecal microbiotafrom a subject selected from the group consisting of a human, a bovine,a dairy calf, a ruminant, an ovine, a caprine, or a cervine. In anotheraspect, a therapeutic composition can be administered to a subjectselected from the group consisting of a human, a bovine, a dairy calf, aruminant, an ovine, a caprine, or a cervine. In an aspect, a therapeuticcomposition is substantially or nearly odourless.

In an aspect, a therapeutic composition provided or administered hereincomprises a fecal microbiota comprising a Shannon Diversity Index ofgreater than or equal to 0.3, greater than or equal to 0.4, greater thanor equal to 0.5, greater than or equal to 0.6, greater than or equal to0.7, greater than or equal to 0.8, greater than or equal to 0.9, greaterthan or equal to 1.0, greater than or equal to 1.1, greater than orequal to 1.2, greater than or equal to 1.3, greater than or equal to1.4, greater than or equal to 1.5, greater than or equal to 1.6, greaterthan or equal to 1.7, greater than or equal to 1.8, greater than orequal to 1.9, greater than or equal to 2.0, greater than or equal to2.1, greater than or equal to 2.2, greater than or equal to 2.3, greaterthan or equal to 2.4, greater than or equal to 2.5, greater than orequal to 3.0, greater than or equal to 3.1, greater than or equal to3.2, greater than or equal to 3.3, greater than or equal to 3.4, greaterthan or equal to 3.5, greater than or equal to 3.6, greater than orequal to 3.7, greater than or equal to 3.8, greater than or equal to3.9, greater than or equal to 4.0, greater than or equal to 4.1, greaterthan or equal to 4.2, greater than or equal to 4.3, greater than orequal to 4.4, greater than or equal to 4.5, or greater than or equal to5.0. In another aspect, a therapeutic composition comprises fecalmicrobiota comprising a Shannon Diversity Index of between 0.1 and 3.0,between 0.1 and 2.5, between 0.1 and 2.4, between 0.1 and 2.3, between0.1 and 2.2, between 0.1 and 2.1, between 0.1 and 2.0, between 0.4 and2.5, between 0.4 and 3.0, between 0.5 and 5.0, between 0.7 and 5.0,between 0.9 and 5.0, between 1.1 and 5.0, between 1.3 and 5.0, between1.5 and 5.0, between 1.7 and 5.0, between 1.9 and 5.0, between 2.1 and5.0, between 2.3 and 5.0, between 2.5 and 5.0, between 2.7 and 5.0,between 2.9 and 5.0, between 3.1 and 5.0, between 3.3 and 5.0, between3.5 and 5.0, between 3.7 and 5.0, between 31.9 and 5.0, or between 4.1and 5.0. In one aspect, a Shannon Diversity Index is calculated at thephylum level. In another aspect, a Shannon Diversity Index is calculatedat the family level. In one aspect, a Shannon Diversity Index iscalculated at the genus level. In another aspect, a Shannon DiversityIndex is calculated at the species level. In a further aspect, atherapeutic composition comprises a preparation of flora in proportionalcontent that resembles a normal healthy human fecal flora.

In a further aspect, a therapeutic composition comprises fecal bacteriafrom at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 different families. Inanother aspect, a therapeutic composition comprises fecal bacteria fromat least 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 different families.In yet another aspect, a therapeutic composition comprises fecalbacteria from at least 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30different families. In a further aspect, a therapeutic compositioncomprises fecal bacteria from at least 31, 32, 33, 34, 35, 36, 37, 38,39, or 40 different families. In another aspect, a therapeuticcomposition comprises fecal bacteria from at least 41, 42, 43, 44, 45,46, 47, 48, 49, or 50 different families. In another aspect, atherapeutic composition comprises fecal bacteria from between 1 and 10,between 10 and 20, between 20 and 30, between 30 and 40, between 40 and50 different families. In an aspect, a therapeutic composition providedor administered herein comprises a fecal microbiota comprising nogreater than 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%,0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% weight non-livingmaterial/weight biological material. In another aspect, a therapeuticcomposition provided or administered herein comprises a fecal microbiotacomprising no greater than 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 75%, 80%, 85%, 90%, or 95% weight non-living material/weightbiological material. In another aspect, a therapeutic compositionprovided or administered herein comprises, consists of, or consistsessentially of, particles of non-living material and/or particles ofbiological material of a fecal sample that passes through a sieve, acolumn, or a similar filtering device having a sieve, exclusion, orparticle filter size of 2.0 mm, 1.0 mm, 0.5 mm, 0.33 mm, 0.25 mm, 0.212mm, 0.180 mm, 0.150 mm, 0.125 mm, 0.106 mm, 0.090 mm, 0.075 mm, 0.063mm, 0.053 mm, 0.045 mm, 0.038 mm, 0.032 mm, 0.025 mm, 0.020 mm, 0.01 mm,or 0.002 mm. “Non-living material” does not include an excipient, e.g.,a pharmaceutically inactive substance, such as a cryoprotectant, addedto a processed fecal material. “Biological material” refers to theliving material in fecal material, and includes microbes includingprokaryotic cells, such as bacteria and archaea (e.g., livingprokaryotic cells and spores that can sporulate to become livingprokaryotic cells), eukaryotic cells such as protozoa and fungi, andviruses. In one embodiment, “biological material” refers to the livingmaterial, e.g., the microbes, eukaryotic cells, and viruses, which arepresent in the colon of a normal healthy human. In an aspect, atherapeutic composition provided or administered herein comprises anextract of human feces where the composition is substantially odorless.In an aspect, a therapeutic composition provided or administered hereincomprises fecal material or a fecal floral preparation in a lyophilized,crude, semi-purified or purified formulation.

In an aspect, a fecal microbiota in a therapeutic composition compriseshighly refined or purified fecal flora, e.g., substantially free ofnon-floral fecal material. In an aspect, a fecal microbiota can befurther processed, e.g., to undergo microfiltration before, after, orbefore and after sieving. In another aspect, a highly purified fecalmicrobiota product is ultra-filtrated to remove large molecules butretain the therapeutic microflora, e.g., bacteria.

In another aspect, a fecal microbiota in a therapeutic composition usedherein comprises or consists essentially of a substantially isolated ora purified fecal flora or entire (or substantially entire) microbiotathat is (or comprises) an isolate of fecal flora that is at least about90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%,99.8% or 99.9% isolated or pure, or having no more than about 0.1%,0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1.0% or more non-fecalfloral material; or, a substantially isolated, purified, orsubstantially entire microbiota as described in Sadowsky et al., WO2012/122478 A1, or as described in Borody et al., WO 2012/016287 A2.

In an aspect, a fecal microbiota in a therapeutic composition comprisesa donor's substantially entire or non-selected fecal microbiota,reconstituted fecal material, or synthetic fecal material. In anotheraspect, the fecal microbiota in a therapeutic composition comprises noantibiotic resistant population. In another aspect, a therapeuticcomposition comprises a fecal microbiota and is largely free ofextraneous matter (e.g., non-living matter including acellular mattersuch as residual fiber, DNA, RNA, viral coat material, non-viablematerial; and living matter such as eukaryotic cells from the fecalmatter's donor).

In an aspect, a fecal microbiota in a therapeutic composition usedherein is derived from disease-screened fresh homologous feces orequivalent freeze-dried and reconstituted feces. In an aspect, a freshhomologous feces does not include an antibiotic resistant population. Inanother aspect, a fecal microbiota in a therapeutic composition isderived from a synthetic fecal composition. In an aspect, a syntheticfecal composition comprises a preparation of viable flora whichpreferably in proportional content, resembles normal healthy human fecalflora which does not include antibiotic resistant populations. Suitablemicroorganisms may be selected from the following: Bacteroides,Eubacterium, Fusobacterium, Propionibacterium, Lactobacillus,Ruminococcus, Escherichia coli, Gemmiger, Clostridium, Desulfomonas,Peptostreptococcus, Bifidobacterium, Collinsella, Coprococcus, Dorea,and Ruminococcus.

In an aspect, a therapeutic composition used in a treatment disclosedherein comprises a sterile fecal filtrate or a non-cellular fecalfiltrate. In one aspect, a sterile fecal filtrate originates from adonor stool. In another aspect, a sterile fecal filtrate originates fromcultured microorganisms. In another aspect, a sterile fecal filtratecomprises a non-cellular non-particulate fecal component. In one aspect,a sterile fecal filtrate is made as described in WO2014/078911,published May 30, 2014. In another aspect, a sterile fecal filtrate ismade as described in Ott et al., Gastroenterology 152:799-911(2017).

In one aspect, a fecal filtrate comprises secreted, execreted orotherwise liquid components or a microbiota, e.g., biologically activemolecules (BAMs), which can be antibiotics or anti-inflammatories, arepreserved, retained or reconstituted in a flora extract.

In one aspect, an exemplary therapeutic composition comprises startingmaterial from a donor from a defined donor pool, where this donorcontributes a stool that is centrifuged, then filtered with veryhigh-level filtration using e.g., either metal sieving or Milliporefilters, or equivalent, to ultimately permit only cells of bacterialorigin to remain, e.g., often less than about 5 micrometers diameter.After the initial centrifugation, the solid material is separated fromthe liquid, and the solid is then filtered in progressively reducingsize filters and tangential filters, e.g., using a Millipore filtration,and optionally, also comprising use of nano-membrane filtering. Thefiltering can also be done by sieves as described in WO 2012/122478, butin contrast using sieves that are smaller than 0.0120 mm, down to about0.0110 mm, which ultimately result in having only bacterial cellspresent.

The supernatant separated during centrifugation is now taken andfiltered progressively in a filtering, e.g., a Millipore filtering orequivalent systems, to end up with liquid which is finely filteredthrough an about 0.22 micron filter. This removes all particulate matterincluding all living matter, including bacteria and viruses. The productthen is sterile, but the aim is to remove the bacteria but to keep theirsecretions, especially antimicrobial bacteriocins, bacteria-derivedcytokine-like products and all accompanying Biologically ActiveMolecules (BAMs), including: thuricin (which is secreted by bacilli indonor stools), bacteriocins (including colicin, troudulixine orputaindicine, or microcin or subtilosin A), lanbiotics (including nisin,subtilin, epidermin, mutacin, mersacidin, actagardine, cinnamycin),lacticins and other antimicrobial or anti-inflammatory compounds.

In one aspect, a therapeutic composition used here comprises areconstituted fecal flora consisting essentially of a combination of apurified fecal microbiota and a non-cellular fecal filtrate. In anotheraspect, a therapeutic composition used here comprises a purified fecalmicrobiota supplemented with one or more non-cellular non-particulatefecal components. In one aspect, a therapeutic composition used herecomprises one or more non-cellular non-particulate fecal components. Inone aspect, one or more non-cellular non-particulate fecal componentscomprise synthetic molecules, biologically active molecules produced bya fecal microorganism, or both. In another aspect, one or morenon-cellular non-particulate fecal components comprise biologicallyactive proteins or peptides, micronutrients, fats, sugars, smallcarbohydrates, trace elements, mineral salts, ash, mucous, amino acids,nutrients, vitamins, minerals, or any combination thereof. In oneaspect, one or more non-cellular non-particulate fecal componentscomprise one or more biologically active molecules selected from thegroup consisting of bacteriocin, lanbiotic, and lacticin. In anotheraspect, one or more non-cellular non-particulate fecal componentscomprise one or more bacteriocins selected from the group consisting ofcolicin, troudulixine, putaindicine, microcin, and subtilosin A. In oneaspect, one or more non-cellular non-particulate fecal componentscomprise one or more lanbiotics selected from the group consisting ofthuricin, nisin, subtilin, epidermin, mutacin, mersacidin, actagardine,and cinnamycin. In another aspect, one or more non-cellularnon-particulate fecal components comprise an anti-spore compound, anantimicrobial compound, an anti-inflammatory compound, or anycombination thereof. In a further aspect, one or more non-cellularnon-particulate fecal components comprise an interleukin, a cytokine, aleukotriene, an eicosanoid, or any combination thereof.

In another aspect, a treatment method provided here comprises the use ofboth fecal bacterial cells, e.g., a partial or a complete representationof the human GI microbiota, and an isolated, processed, filtered,concentrated, reconstituted and/or artificial liquid component (e.g.,fecal filtrate) of the flora (the microbiota) which comprises, amongothers ingredients, bacterial secretory products such as e.g.,bacteriocins (proteinaceous toxins produced by bacteria, includingcolicin, troudulixine or putaindicine, or microcin or subtilosin A),lanbiotics (a class of peptide antibiotics that contain a characteristicpolycyclic thioether amino acid lanthionine or methyllanthionine, andunsaturated amino acids dehydroalanine and 2-aminoisobutyric acid; whichinclude thuricin (which is secreted by bacilli in donor stools), nisin,subtilin, epidermin, mutacin, mersacidin, actagardine, cinnamycin), alacticin (a family of pore-forming peptidic toxins) and otherantimicrobial or anti-inflammatory compounds and/or additionalbiologically active molecules (BAMs) produced by bacteria or othermicroorganisms of the microbiota, and/or which are found in the “liquidcomponent” of a microbiota.

In one aspect, a fecal bacteria-based therapeutic composition is usedconcurrently with a fecal non-cellular filtrate-based therapeuticcomposition. In another aspect, a patient is treated with a first fecalnon-cellular filtrate-based therapeutic composition before being given asecond fecal bacteria-based therapeutic composition, or vice versa. In afurther aspect, a treatment method comprises three steps: first,antibiotic pre-treatment to non-selectively remove infectiouspathogen(s); second, a fecal non-cellular filtrate-based treatment stepto further suppress selected infectious pathogen(s); and third, givingthe patient a fecal bacteria-based therapeutic composition tore-establish a functional intestinal microbiome.

In an aspect, a therapeutic composition is combined with other adjuvantssuch as antacids to dampen bacterial inactivation in the stomach. (e.g.,Mylanta, Mucaine, Gastrogel). In another aspect, acid secretion in thestomach could also be pharmacologically suppressed using H2-antagonistsor proton pump inhibitors. An example H2-antagonist is ranitidine. Anexample proton pump inhibitor is omeprazole. In one aspect, an acidsuppressant is administered prior to administering, or inco-administration with, a therapeutic composition.

In an aspect, a therapeutic composition is in the form of: an enemacomposition which can be reconstituted with an appropriate diluent;enteric-coated capsules; enteric-coated microcapsules; acid-resistanttablet; acid-resistant capsules; acid-resistant microcapsules; powderfor reconstitution with an appropriate diluent for naso-enteric infusionor colonoscopic infusion; powder for reconstitution with appropriatediluent, flavoring and gastric acid suppression agent for oralingestion; powder for reconstitution with food or drink; or food or foodsupplement comprising enteric-coated and/or acid-resistant microcapsulesof the composition, powder, jelly, or liquid.

In an aspect, a treatment method effects a cure, reduction of thesymptoms, or a percentage reduction of symptoms of multiple sclerosis.The change of flora is preferably as “near-complete” as possible and theflora is replaced by viable organisms which will crowd out anyremaining, original flora. Typically the change in enteric floracomprises introduction of an array of predetermined flora into thegastro-intestinal system, and thus in a preferred form the method oftreatment comprises substantially or completely displacing pathogenicenteric flora in patients requiring such treatment.

In another aspect, a therapeutic composition can be provided togetherwith a pharmaceutically acceptable carrier. As used herein, a“pharmaceutically acceptable carrier” refers to a non-toxic solvent,dispersant, excipient, adjuvant, or other material which is mixed with alive bacterium in order to permit the formation of a pharmaceuticalcomposition, e.g., a dosage form capable of administration to thepatient. A pharmaceutically acceptable carrier can be liquid (e.g.,saline), gel or solid form of diluents, adjuvant, excipients or an acidresistant encapsulated ingredient. Suitable diluents and excipientsinclude pharmaceutical grades of physiological saline, dextrose,glycerol, mannitol, lactose, starch, magnesium stearate, sodiumsaccharin, cellulose, magnesium carbonate, and the like, andcombinations thereof. In another aspect, a therapeutic composition maycontain auxiliary substances such as wetting or emulsifying agents,stabilizing or pH buffering agents. In an aspect, a therapeuticcomposition contains about 1%-5%, 5%-10%, 10%-15%, 15-20%, 20%-25%,25-30%, 30-35%, 40-45%, 50%-55%, 1%-95%, 2%-95%, 5%-95%, 10%-95%,15%-95%, 20%-95%, 25%-95%, 30%-95%, 35%-95%, 40%-95%, 45%-95%, 50%-95%,55%-95%, 60%-95%, 65%-95%, 70%-95%, 45%-95%, 80%-95%, or 85%-95% ofactive ingredient. In an aspect, a therapeutic composition containsabout 2%-70%, 5%-60%, 10%-50%, 15%-40%, 20%-30%, 25%-60%, 30%-60%, or35%-60% of active ingredient.

In an aspect, a therapeutic composition can be incorporated intotablets, drenches, boluses, capsules or premixes. Formulation of theseactive ingredients into such dosage forms can be accomplished by meansof methods well known in the pharmaceutical formulation arts. See, e.g.,U.S. Pat. No. 4,394,377. Filling gelatin capsules with any desired formof the active ingredients readily produces capsules. If desired, thesematerials can be diluted with an inert powdered diluent, such as sugar,starch, powdered milk, purified crystalline cellulose, or the like toincrease the volume for convenience of filling capsules.

In an aspect, conventional formulation processes can be used to preparetablets containing a therapeutic composition. In addition to the activeingredients, tablets may contain a base, a disintegrator, an absorbent,a binder, and a lubricant. Typical bases include lactose, sugar, sodiumchloride, starch and mannitol. Starch is also a good disintegrator as isalginic acid. Surface-active agents such as sodium lauryl sulfate anddioctyl sodium sulphosuccinate are also sometimes used. Commonly usedabsorbents include starch and lactose. Magnesium carbonate is alsouseful for oily substances. As a binder there can be used, for example,gelatin, gums, starch, dextrin, polyvinyl pyrrolidone and variouscellulose derivatives. Among the commonly used lubricants are magnesiumstearate, talc, paraffin wax, various metallic soaps, and polyethyleneglycol.

In an aspect, for preparing solid compositions such as tablets, anactive ingredient is mixed with a pharmaceutical carrier, e.g.,conventional tableting ingredients such as corn starch, lactose,sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalciumphosphate or gums, or other pharmaceutical diluents, e.g. water, to forma solid preformulation composition containing a homogeneous mixture of acomposition of the present invention. When referring to thesepreformulation compositions as homogeneous, it is meant that the activeingredient is dispersed evenly throughout the composition so that thecomposition may be readily subdivided into equally effective unit dosageforms such as tablets, pills and capsules. This solid preformulationcomposition is then subdivided into unit dosage forms of the typedescribed above containing a desired amount of an active ingredient(e.g., at least about 10⁵, 10⁶, 10⁷, 10⁸, 10⁹, 10¹⁰, 10¹¹, 10¹², or 10¹³cfu). A therapeutic composition used herein can be flavored.

In an aspect, a therapeutic composition can be a tablet or a pill. Inone aspect, a tablet or a pill can be coated or otherwise compounded toprovide a dosage form affording the advantage of prolonged action. Forexample, a tablet or pill can comprise an inner dosage and an outerdosage component, the latter being in the form of an envelope over theformer. The two components can be separated by an enteric layer whichserves to resist disintegration in the stomach and permits the innercomponent to pass intact into the duodenum or to be delayed in release.A variety of materials can be used for such enteric layers or coatings,such materials including a number of polymeric acids and mixtures ofpolymeric acids with such materials as shellac, cetyl alcohol andcellulose acetate.

In an aspect, a therapeutic composition can be a drench. In one aspect,a drench is prepared by choosing a saline-suspended form of atherapeutic composition. A water-soluble form of one ingredient can beused in conjunction with a water-insoluble form of the other bypreparing a suspension of one with an aqueous solution of the other.Water-insoluble forms of either active ingredient may be prepared as asuspension or in some physiologically acceptable solvent such aspolyethylene glycol. Suspensions of water-insoluble forms of eitheractive ingredient can be prepared in oils such as peanut, corn, sesameoil or the like; in a glycol such as propylene glycol or a polyethyleneglycol; or in water depending on the solubility of a particular activeingredient. Suitable physiologically acceptable adjuvants may benecessary in order to keep the active ingredients suspended. Adjuvantscan include and be chosen from among the thickeners, such ascarboxymethylcellulose, polyvinyl pyrrolidone, gelatin and thealginates. Surfactants generally will serve to suspend the activeingredients, particularly the fat-soluble propionate-enhancingcompounds. Most useful for making suspensions in liquid nonsolvents arealkylphenol polyethylene oxide adducts, naphthalenesulfonates,alkylbenzene-sulfonates, and the polyoxyethylene sorbitan esters. Inaddition many substances, which affect the hydrophilicity, density andsurface tension of the liquid, can assist in making suspensions inindividual cases. For example, silicone anti-foams, glycols, sorbitol,and sugars can be useful suspending agents.

In an aspect, a therapeutic composition comprises non-pathogenic sporesof one or more, two or more, three or more, or four or more Clostridiumspecies selected from the group consisting of Clostridium absonum,Clostridium argentinense, Clostridium baratii, Clostridium botulinum,Clostridium cadaveris, Clostridium carnis, Clostridium celatum,Clostridium chauvoei, Clostridium clostridioforme, Clostridiumcochlearium, Clostridium fallax, Clostridium felsineum, Clostridiumghonii, Clostridium glycolicum, Clostridium haemolyticum, Clostridiumhastiforme, Clostridium histolyticum, Clostridium indolis, Clostridiumirregulare, Clostridium limosum, Clostridium malenominatum, Clostridiumnovyi, Clostridium oroticum, Clostridium paraputrificum, Clostridiumperfringens, Clostridium piliforme, Clostridium putrefaciens,Clostridium putrificum, Clostridium sardiniense, Clostridiumsartagoforme, Clostridium scindens, Clostridium septicum, Clostridiumsordellii, Clostridium sphenoides, Clostridium spiroforme, Clostridiumsporogenes, Clostridium subterminale, Clostridium symbiosum, Clostridiumtertium, Clostridium tetani, Clostridium welchii, and Clostridiumvillosum.

In an aspect, a therapeutic composition comprises purified, isolated, orcultured viable non-pathogenic Clostridium and a plurality of purified,isolated, or cultured viable non-pathogenic microorganisms from one ormore genera selected from the group consisting of Collinsella,Coprococcus, Dorea, Eubacterium, and Ruminococcus. In another aspect, atherapeutic composition comprises a plurality of purified, isolated, orcultured viable non-pathogenic microorganisms from one or more generaselected from the group consisting of Clostridium, Collinsella,Coprococcus, Dorea, Eubacterium, and Ruminococcus.

In an aspect, a therapeutic composition comprises two or more generaselected from the group consisting of Collinsella, Coprococcus, Dorea,Eubacterium, and Ruminococcus. In another aspect, a therapeuticcomposition comprises two or more genera selected from the groupconsisting of Coprococcus, Dorea, Eubacterium, and Ruminococcus. In afurther aspect, a therapeutic composition comprises one or more, two ormore, three or more, four or more, or five or more species selected fromthe group consisting of Coprococcus catus, Coprococcus comes, Dorealongicatena, Eubacterium eligens, Eubacterium hadrum, Eubacteriumhallii, Eubacterium rectale, and Ruminococcus torques.

In one aspect, a therapeutic composition comprises at least about 10⁵,10⁶, 10⁷, 10⁸, 10⁹, 10¹⁰, 10¹¹, 10¹², or 10¹³ cfu or total cell count.In another aspect, a therapeutic composition comprises at most about10⁵, 10⁶, 10⁷, 10⁸, 10⁹, 10¹⁰, 10¹¹, 10¹², 10¹³ or 10¹⁴ cells or totalcell count.

In another aspect, a therapeutic composition comprises at least about10⁵, 10⁶, 10⁷, 10⁸, 10⁹, 10¹⁰, 10¹¹, 10¹², or 10¹³ cells or total cellcount. In another aspect, a therapeutic composition comprises at mostabout 10⁵, 10⁶, 10⁷, 10⁸, 10⁹, 10¹⁰, 10¹¹, 10¹², 10¹³ or 10¹⁴ cells ortotal cell count.

In one aspect, a therapeutic composition is formulated as an oralcapsule, microcapsule, tablet, or pill. In another aspect, a capsule,microcapsule, tablet, or pill is adapted for enteric delivery. In afurther aspect, a capsule, microcapsule, tablet, or pill is an entericcapsule, microcapsule, tablet, or pill. In another aspect, a capsule,microcapsule, tablet, or pill comprises an enteric coating, is acidresistant, or both.

EXAMPLES Example 1 Preparation of Fecal Microbiota

Fecal microbiota is prepared essentially according to protocolspublished in US2014/0147417 or WO2014/152484. Summarized below is anexemplary protocol.

Potential fecal microbiota donors are screened according to a list ofcriteria used to exclude unsuitable donors. Potential fecal microbiotadonors are excluded if they have received antibiotics, laxatives, dietpills, immunomodulators or chemotherapy in the preceding three months.Potential fecal microbiota donors are excluded if they have a history ofall known infectious diseases, morbid obesity, diabetes, irritable bowelsyndrome, inflammatory bowel disease, chronic diarrhea, constipation,colorectal polyps or cancer, a compromised immune system, metabolicsyndromes, chronic fatigue syndrome, major GI surgery, or other diseasesor conditions potentially associated with specific changes in fecalmicrobiota. Potential fecal microbiota donors are excluded if theyexhibit positive laboratory tests for C-reactive protein, erythrocytesedimentation rate, hepatitis A, hepatitis B, hepatitis C, humanimmunodeficiency virus, human T-lymphotropic virus, or syphilis.Potential fecal microbiota donors are excluded if they exhibit apositive test for stool ova, parasites, and/or viruses. Potential fecalmicrobiota donors are excluded if they engage in high-risk sexualbehaviors, have been incarcerated, or received any tattoos or bodypiercings in areas that have had disease epidemics within the past threemonths.

Donor fecal material (fresh feces) is collected in a sterilizedcontainer, and then it is transferred to a blender. Approximately500-1000 mL 0.9% saline solution is added to the blender and thoroughlymixed with the fecal sample. The resulting suspension is filtered atleast 4 times through strainers prior to collecting a final suspension.The final suspension is centrifuged in 50 mL tubes at 1200×g for 3minutes. The supernatant is discarded and the pellet is gentlyresuspended in approximately 50 mL of sterile 0.9% saline solution. Thecentrifugation and resuspension steps are repeated 2 to 4 additionaltimes. Upon the final centrifugation, the supernatant is discarded. Ifthe fecal microbiota is to be used immediately, the resultant pellet isresuspended in 1.5-volumes of 0.9% saline solution by gently mixing. Ifthe fecal microbiota is to be stored, the resultant pellet isresuspended in 10% sterile glycerol and stored at −80 degreesCentigrade. If fecal microbiota are frozen, they are warmed to roomtemperature prior to administration to a patient

Example 2 Treatment of Multiple Sclerosis Induced Constipation inWheelchair-Bound Patients

A 30-year old male patient presents with constipation and a recenthistory of multiple sclerosis. The patient suddenly develops symptoms ofvertigo, impaired concentration and mood alterations post a surgery. Thesymptoms further lead to neuritis and trigeminal neuralgia and result ina multiple sclerosis diagnosis which was confirmed by MRI. Prior to thefecal microbiome therapy, the patient receives interferon treatmentswhich do not improve his condition. Fecal microbiome therapy isadministered for constipation, resulting in complete resolution. Thepatient's MS symptoms progressively improve and the ability to walk isregained, facilitating catheter removal. The patient remainsasymptomatic 15-years post-fecal microbiome therapy without relapse.Follow-up MRI 15-years post-fecal microbiome therapy reports a haltingof disease progression and no evidence of active disease.

A 29-year old male presents with severe, chronic constipation, andatypical multiple sclerosis. Bilateral leg paralysis and urinaryincontinence leaves the patient wheelchair-bound with a indwellingurinary catheter. Fecal microbiome therapy results in dramaticresolution of constipation, allowing daily defecation with ease. Thepatient also reports a rapid and progressive improvement in multiplesclerosis symptoms. Improvements include the patient regaining theability to walk, defecate and urinate normally, and subsequent urinarycatheter removal. The patient remains asymptomatic 7-years post-fecalmicrobiome therapy.

An 80-year old female patient presents with severe chronic constipation,and a history of atypical multiple sclerosis manifesting as a severemuscular weakness, leaving the patient wheelchair-bound. Fecalmicrobiome therapy treatment results in rapid improvement ofconstipation and increased energy levels. At eight months post-Fecalmicrobiome therapy the patient reports complete resolution of bowelsymptoms and neurological improvement. The patient gains the ability towalk long distances unassisted. Two years post-Fecal microbiome therapy,the patient remains asymptomatic.

Example 3 Treatment of Multiple Sclerosis and Irritable Bowel Syndrome(IBS)

A 39-year old male patient, with longstanding irritable bowel syndromeand Multiple Sclerosis, that has been dormant and stable for two years,is treated with Fecal microbiome therapy. The patient's symptoms includemuscle fatigue, loss of balance, walking difficulty with much weaknessand required support, and slurred speech. The patient also showsdemyelination on MRI which later improves on a repeat MRI.

The patient is treated with fecal microbiome therapy through thecolonoscope into the ascending colon, followed by nine enemas over thenext week to increase the bacterial load. The patient initially respondsfor two months and shows no symptoms of either IBS or MS. The patientregains energy and has no feelings of nausea. Further, the patientpasses two formed motions per day, and he is able to walk withoutsupport after treatment. This marks the only period of good healthexperienced by the patient over many years. However, after two months ofimprovement, his symptoms started recurring and he reverted to where hiscolonic flora was in the first place. The patient was prescribedRifaximin and low dose probiotics, which had little effect on hismultiple sclerosis and IBS symptoms.

Example 4 Treatment of Multiple Sclerosis

A 57-year old male patient with multiple sclerosis is treated with fecalmicrobiome therapy. The patient suffers from lethargy, weakness, weightloss, altered bowel habits, and urinary incontinence. Three months afterFecal microbiome therapy treatment, the patient experiences increasedenergy and his constipation is deemed to be resolved. However, eightmonths after his fecal microbiome therapy treatment the initial symptomsreoccur. The patient begins a course of antibiotic pre-treatment inpreparation for a repeat colonic fecal microbiome therapy.

Additional treatments followed by home infusions to continue thepreviously gained improvements are performed.

Example 5 Treatment of Multiple Sclerosis

A female patient, suffering from multiple sclerosis and constipation isprescribed fecal microbiome therapy pre-treatment consisting ofRifaximin and Vancomycin. The pre-treatment improves the patient'smultiple sclerosis symptoms and the patient experiences resolution ofconstipation, increased energy, and ability to move affected musclesmore strongly. Following pre-treatment and cessation of antibiotics, thepatient receives fecal microbiome therapy. Approximately one and a halfmonths after the fecal microbiome therapy, the patient reportsexperiencing an improvement in energy, decreased bloating andincontinence, and improved balance. The patient continues receivingfecal microbiome therapy to treat gastrointestinal symptoms and hermultiple sclerosis attributed weakness. In summary, fecal microbiometherapy improves the patient's gastrointestinal symptoms and weakness asa result of multiple sclerosis.

Example 6 Treatment of Multiple Sclerosis

A 27 year old patient, suffers from multiple sclerosis, a stiff and weakright leg, a 4/5 weakness for right hip flexion, a 3-4/5 weak kneeflexion, a 4+/5 ankle dorsiflexion, and 4+/5 weakness for right handabduction weakness in right shoulder. The patient is prescribed fecalmicrobiome therapy.

Example 7 Oral Capsule Treatment Protocol for Multiple Sclerosis

Patients are divided into four groups (Groups 1 to 4). Group 1 patientsare administered a pre-treatment of antibiotics (e.g., Vancomycin andMetronidazole). Group 2 receives no antibiotics. Both Groups 1 and 2receive a pre-colonoscopy bowel prep followed by capsule fecalmicrobiome therapy. Groups 3 and 4 receive no bowel prep while Group 3,but not Group 4, also receives an antibiotic pretreatment. A singlecapsule contains between 10⁹ and 10¹² viable cells. Capsules areadministered for 18 weeks as follows: two capsules twice-a-day for 14days, two capsules twice-a-day every other day for 14 days, 4 capsulestwice-a-week for 14 days, and 4 capsules once-a-week (e.g., each Monday)for 12 weeks. High dose capsules (total cell count of about 10¹²) areused in loading doses (also called treatment doses) for the initial 4weeks. Lower dose capsules (total cell count of about 10⁹) are used inmaintenance doses for the subsequent 14 weeks. In patients receivingantibiotic pretreatment, capsules are administered one day after ceasingantibiotics. Patient symptoms are observed and clinical examination isperformed before, during and post oral capsule treatment. Pre, duringand post-treatment DNA metagenomics (2-4 days; 1 week; 6 weeks; 12weeks) are also carried out. The capsule treatments reverse patientsymptoms and result in a clinically normal urge and defecation.

As various modifications could be made in the constructions and methodsherein described and illustrated without departing from the scope of thedisclosure, it is intended that all matter contained in the foregoingdescription shall be interpreted as illustrative rather than limiting.The breadth and scope of the present disclosure should not be limited byany of the above-described exemplary embodiments, but should be definedonly in accordance with the following claims appended hereto and theirequivalents. All patent and non-patent documents cited in thisspecification are incorporated herein by reference in their entirety.

1. A method for treating multiple sclerosis in a human subject in needthereof, said method comprising orally administering to said humansubject a pharmaceutically active dose of a therapeutic compositioncomprising live non-pathogenic fecal bacteria. 2.-114. (canceled) 115.The method of claim 1, wherein said multiple sclerosis is selected fromthe group consisting of clinically isolated syndrome (CIS),relapsing-remitting multiple sclerosis (RRMS), primary progressivemultiple sclerosis (PPMS), and secondary progressive multiple sclerosis(SPMS).
 116. The method of claim 1, wherein said therapeutic compositioncomprises a non-selected fecal microbiota.
 117. The method of claim 116,wherein said fecal microbiota comprises a donor's entire fecalmicrobiota.
 118. The method of claim 116, wherein said fecal microbiotais from synthetic fecal material.
 119. The method of claim 1, whereinsaid method reduces the Sum8 score of said subject by at least 10%, 20%,30%, 50%, 60%, 70%, 80%, or 90% after at least 8 weeks of treatment.120. The method of claim 1, wherein said dose is administered at leastonce daily for at least 2 weeks.
 121. The method of claim 1, whereinsaid dose is administered at least twice weekly for at least two weeks.122. The method of claim 121, wherein said dose is administered at leasttwice weekly for at least 4 weeks.
 123. The method of claim 121, whereinsaid dose is administered at least twice weekly for at least 8 weeks.124. The method of claim 1, wherein said dose is administered at leastthree times weekly for at least 4 weeks.
 125. The method of claim 1,wherein said therapeutic composition comprises both live non-pathogenicfecal bacteria and a non-cellular fecal filtrate.
 126. The method ofclaim 1, wherein said method comprises a first dosing schedule followedby a second dosing schedule, wherein said second dosing schedulecomprises a maintenance dose lower or equal to the dose of said firstdosing schedule.
 127. The method of claim 1, wherein said therapeuticcomposition is formulated as a delayed or gradual enteric release farm.128. The method of claim 1, wherein said therapeutic composition isformulated as an enteric coated capsule or an acid-resistant capsule.129. The method of claim 1, wherein said pharmaceutically active dose isfrom 10⁸ to 10¹⁴ cfu or total number of cells.
 130. The method of claim1, wherein said therapeutic composition is in a liquid, frozen,freeze-dried, spray-dried, foam-dried, or powder form.
 131. The methodof claim 1, wherein said therapeutic composition comprises acryoprotectant selected from the group consisting of polyethyleneglycol, skim milk, erythritol, arabitol, sorbitol, glucose, fructose,alanine, glycine, proline, sucrose, lactose, ribose, trehalose, dimethylsulfoxide (DMSO), glycerol, and a combination thereof.
 132. The methodof claim 1, wherein said subject is pretreated with an antibiotic priorto administration of said composition.
 133. The method of claim 1,wherein said method eliminates or reduces one or more symptoms selectedfrom the group consisting of constipation and stool leakage, difficultyurinating, frequent urination, urine leakage, double vision, eyediscomfort, rapid eye movements, vision loss, memory loss, difficultysolving problems, depression, hearing loss, slurred speech, difficultywith chewing and difficulty with swallowing, limb weakness, tremors,numbness, and motor difficulties.